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Phony Phone

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Agar transfer, avoiding isolation
#27001555 - 10/24/20 04:44 PM (3 years, 3 months ago) |
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Hello shroomery, sorry if this has been asked already. I wanted to ask, if you wanted to transfer a clone mycelium from plate #1 to plate #2 but you wanted to avoid further isolating the strains, is the solution to this to just grab a big enough wedge compared to a tiny one?
Thanks for your time.
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Lenz
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Re: Agar transfer, avoiding isolation [Re: Phony Phone]
#27001633 - 10/24/20 05:41 PM (3 years, 3 months ago) |
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I was wondering about this myself recently cause I've been taking a bunch of transfers from clone plates.
Here's a good thread about it: https://www.shroomery.org/forums/showflat.php/Number/23660093#23660093
Quote:
Homesteader said: Hey morty great post and you are definitely on the right track brother! This topic is one I struggled with most when starting on Agar. What I've noticed is that a clone is usually comprised of multiple isolates fused together through anastamosis. They will hold together as one uniform 'type' of mycelium for several transfers. However, 3-5 transfers (Increase in P value) later I notice sectoring of once stable strains. Transferring these sectors will leave you with an isolate that no longer contains all the genetic material that once made up your cloned fruit and may no longer even have the capacity to fruit. To combat this I keep as low P value as possible and avoid unnecessary transfers. I also change up the nutrient concentrations of my Agar blend and sometimes alternate the actual source of carbohydrates (one time PDA, next time MEA).
The advantage to mycelium of a cloned fruit is the fact that it is an exact replica of the parent. The disadvantage here (so I'm told) is that some or many of the isolates that make up the cloned fruit may in fact only have the ability to grow vegetatively and have no ability to fruit. Thus, robbing nutrients from the isolates that do have the ability to fruit. This should make no difference whatsoever as the strains that make up a clone are anastomosed and can share their nutrients across the entire network. In theory they work together and not against one another.
To answer your question though: yes you are isolating by trying to clean up your plates and yes you could most certainly end up with an isolate that has no ability to fruit. Is this likely to happen? Not really and it depends on how many times you are transferring. Keep in mind, if you are grabbing rhizomorphic sectors of fast and healthy growing mycelium than you will almost always have a fruiting strain.
Sorry if some of this info is redundant but I felt this is an important topic to cover for people getting started on Agar.
I can't vouch personally for this but a TC in that thread seems to be saying essentially the same thing, that it's not really something to worry about.
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Brian Jones
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Re: Agar transfer, avoiding isolation [Re: Phony Phone]
#27001821 - 10/24/20 07:54 PM (3 years, 3 months ago) |
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It's hard to get isolations even if your trying, taking multiple sequences of transfers from clearly delineated sectors. I would keep transferring small pieces of agar to lower the odds of contamination.
I think your theory was good, but it's not really a problem.
-------------------- "The Rolling Stones will break up over Brian Jones' dead body" John Lennon I don't want no commies in my car. No Christians either. The worst thing about corruption is that it works so well,
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Phony Phone

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Re: Agar transfer, avoiding isolation [Re: Brian Jones]
#27002894 - 10/25/20 02:41 PM (3 years, 3 months ago) |
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thank you both! So it appears that if you were to isolate the strain cluster synergy of the "isolate" clone, u have to go 4-5+ tranfers to start having problems, correct?
some plates might have contam issues so they will need at least 1 more transfer. i am having trouble recognizing the sectors early on as they are so joined up together (the non obvious ones) that I cannot tell if im grabbing a single sectors or multiple "invisible" ones.
How do you guys tell your sectors apart?
Another small question, what does the alternation between agar recipes do for the health of the myc? why is it beneficial to it?
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Big_Dub
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Re: Agar transfer, avoiding isolation [Re: Phony Phone]
#27002958 - 10/25/20 03:07 PM (3 years, 3 months ago) |
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you always want to make tiny transfers. the point of transfering is to get clean healthy mycelium. less is more
look for nice spots, and take those. dont get caught up worrying up sectors. just grow some mushrooms
-------------------- split_by_nine said: click me you fuck do the right thing
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Phony Phone

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Re: Agar transfer, avoiding isolation [Re: Big_Dub]
#27002978 - 10/25/20 03:22 PM (3 years, 3 months ago) |
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with the above information mushrooms might not grow if clones keep getting transferred though 
Quote:
Keep in mind, if you are grabbing rhizomorphic sectors of fast and healthy growing mycelium than you will almost always have a fruiting strain.
why are rhizomorphs certain fruiters?
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Big_Dub
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Re: Agar transfer, avoiding isolation [Re: Phony Phone]
#27002988 - 10/25/20 03:31 PM (3 years, 3 months ago) |
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rhizomorphic mycelium does not equal fruiting. that is a myth. if you want a fruiting culture, wait for pins to show upp an agar, then transfer the pins
-------------------- split_by_nine said: click me you fuck do the right thing
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Phony Phone

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Re: Agar transfer, avoiding isolation [Re: Big_Dub]
#27002997 - 10/25/20 03:37 PM (3 years, 3 months ago) |
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great idea. thanks how do you recognize sectors?
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Big_Dub
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Re: Agar transfer, avoiding isolation [Re: Phony Phone]
#27003002 - 10/25/20 03:45 PM (3 years, 3 months ago) |
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not necessary ime.
just take something from the leading edge. dont overthink this
-------------------- split_by_nine said: click me you fuck do the right thing
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: Phony Phone]
#27003010 - 10/25/20 03:51 PM (3 years, 3 months ago) |
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OP, this is a good question and one that many have thought and asked. As far as I know, no one has a concrete answer. Most of us that have been at it a while were worried about it (as you are) at first but then learned that it doesn't seem to be much of a concern.
I've done lots of tests to try and figure this out and still don't have a perfect answer. I used to always try to take a piece from each obvious sector anytime i was transferring an obviously sectoring clone. I've done a lot of trying one sector vs trying another vs trying all of them together and never came to a solid conclusion. Best i can say is that it's probably not worth tripping on but in theory, you could end up snatching a less optimal set of genes.
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Phony Phone

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Alright, thanks for the tip. Guess keeping the master plates is a fail-safe method to fall back onto if everything else comes out to be bunk.
I have 2 more questions, 1 is, in the refrigerator, how long will cube mycelium live at say around 44 f (6-8 Celsius)?
2, I've read Stro's thread on sectors but I am still kind of confused about it, for example when you have a colony half tomentose - half rhizo on agar, is this sectoring? And on a rhizo growth, are different growth speed patterns obvious sectoring? And within that sector, how do you tell if it's a monoculture sector or just dense strains? is the only way to distinguish that even more transfers until you get the obvious "lines of separation"? Sorry if i got any terminology wrong, feel free to correct me.
Thanks for your time.
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: Phony Phone]
#27003038 - 10/25/20 04:12 PM (3 years, 3 months ago) |
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Properly kept cultures will last a good long while in the fridge. I haven't done exact tests to give an exact answer but I've pulled them out a year later. Thinner plates dry out faster than thicker plates and wrapping them really really well helps. They don't need to breathe in cold storage so you can seal them up nicely.
Slants are what you want for real long term (years) storage.
Sectoring is just what it sounds like. If you can see a difference between this and that it's a "sector". It's not really worth tripping on. The confusion comes when noobs start to think that sectoring = genetic difference absolutely. This is not the case. I have done quite a bit of fooling around with sectors and seen everything and anything possible happen.
I'd love to give you a simple yes or no but.... The best advice is just to transfer from the fastest growing/ most organized growth, do so until the whole culture looks nice and clean, put to grain... to bulk... to fruit. If it kicks ass, keep growing it.
Check my main threads. I did a little write up for some one that i called "masters and slaves...." and i documented a little isolation that shows shitloads of cultures going plate to plate to plate to plate to......
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Phony Phone

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yup, will do. thanks a ton. I got 1 more little thing, what is the theory of alternating between recipes when you transfer myc? What is in that method that help myc from accumulating senescence errors?
Really helpful info
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: Phony Phone]
#27003093 - 10/25/20 04:37 PM (3 years, 3 months ago) |
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The UNPROVEN RUMOR is based on the assumption that the genetics will get so used to surviving off of that type of food that it will get lazy and become a slower growing culture. I have seen no evidence of this in my own life. As a noob i took the rumor seriously and tried to fuck around and find out. I found out that I found no advantage to switching things up and have only and always used Malt extract agar since then.
Others may disagree, I'm just stating what I've seen in my own rainbow farming life.
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Phony Phone

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Say one had only 1 slant and that slant started running out of myc from transfers, would one then take the final bit of myc left in the slant and make a new master slant or a petri? I am afraid this is kind of a silly question but I really wonder what techniques people utilize to keep their favorite cultures running.
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: Phony Phone]
#27003235 - 10/25/20 05:49 PM (3 years, 3 months ago) |
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I'd have a master plate going already and make a new slant from the master plate. If you don't have one, make one so you can verify it's still good and then make another slant.
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NecroMyce
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Quote:
verum subsequentis said: I'd have a master plate going already and make a new slant from the master plate. If you don't have one, make one so you can verify it's still good and then make another slant.
I have been tagging along and I see you post everywhere, Verum. Always helpful bro! Thanks for helping us both out. I have a quick question to chime in, now do You deal with a plate that has a contam in it? Just move slowly and try not to touch the mold or knock the Petri dish hard to avoid disbursing spores everywhere? Here is a few plates with mold or some contam, to give a example
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: NecroMyce]
#27003400 - 10/25/20 06:54 PM (3 years, 3 months ago) |
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You got it. In a still air box in a still room, you gently unwrap (we should talk about your wrapping) the plate. Then flame sterilize the scalpel until red hot outside the box and gently move it into the box. Be sure not to disturb the air in the box in any way. Let you're scalpel cool in the air or cool it quickly in a fresh plate. Every move you make should be gentle and smooth. Once cool reach in and gently take a transfer from as far away from the away from the contam and from the best looking myc around.
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NecroMyce
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I know, I didn’t have any parafilm at the time so I used plastic wrap! Lol and so basically we are looking to just clean up our cultures by doing transfers over time and try to only keep it around 2-3 transfers. What is usually colonization time with agar inoculation?
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: NecroMyce]
#27003624 - 10/25/20 08:40 PM (3 years, 3 months ago) |
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I use glad cling wrap cut into rolls that are just wider than a petri.
2-3 transfers is just a guideline we give people to make sure they know to make sure they've actually achieved a clean plate. It's generally recommended that noobs make one transfer even after they think it's clean. Just because it's better safe than sorry and noobs don't have well trained eyes yet.
Colonization times depend on many factors. Grain prep/ ambient room temp/ how much agar you put in/ how much grain it's trying to colonize/ Vigor of the culture. Two to three weeks is about right though.
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NecroMyce
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Makes sense, thank you. Would you suggest that on your 2nd-3rd transfer, you might want to let it grow all the way to each edge of the plate and fully colonized the plate, maybe even wait for pins to occur.. To make sure no contams pop up?
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: NecroMyce] 1
#27003646 - 10/25/20 08:54 PM (3 years, 3 months ago) |
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Hell no. Contams are generally harder to see on an over colonized plate. Once you have a clean culture (don't think about number of transfer, just transfer until clean) let it grow out 3/4 of the plate and then put to grain. I like to put one plate to four masters but you can stretch it further for sure.
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fightingcherries



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Have you noticed much variation from cloning a mushroom from spawn that was from a clone? My master froze in a shitty fridge, so I had to clone when my tub fruited to try to get my culture back.
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verum subsequentis
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I've only cloned a clone once and it went wonderfully. Never cloned a clone of a clone though. I wouldn't advise against trying it.
You gotta watch out for shitty fridges. Specially ones with freezers in them.
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verum subsequentis
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Also, girlfriends that accidentally leave the door open and cause everything to freeze.
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GrinchGrower
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Quote:
verum subsequentis said: Also, girlfriends that accidentally leave the door open and cause everything to freeze.


Seriously though, thanks for all the good info here for noob agar work verum! Got a couple spore to LME plates of PFC and AC just starting to show some growth that I am excited to work with soon.
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: GrinchGrower]
#27003883 - 10/26/20 12:05 AM (3 years, 3 months ago) |
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My pleasure.
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NecroMyce
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I just made 60 plates, did about 5 transfers of each strain. Tossed on that went too long and had a bunch of contams. So I assume as transfers go on, I’ll have a bunch of masters and I also dropped some fresh fruit to plates. I’m excited to see what’ll come in the future. So say, they grow out to 3/4 the plate, we can put them in the fridge at normal fridge temps which I believe is 34-40 F. And this will stall growth for a while, can you put unused agar plates that are already prepped for storage in the fridge as well? What do you think shelf live is?
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BrotherDekatessera
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Re: Agar transfer, avoiding isolation [Re: Big_Dub]
#27004383 - 10/26/20 10:50 AM (3 years, 3 months ago) |
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Quote:
Big_Dub said: you always want to make tiny transfers. the point of transfering is to get clean healthy mycelium. less is more
look for nice spots, and take those. dont get caught up worrying up sectors. just grow some mushrooms
tbh this is usually the case, just stop worrying and grow more mushrooms.
this latest experiment was the first one where I went simple and easy with transfers, and honestly got the fastest healthiest myc I have seen in a long long time.
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verum subsequentis
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Fresh plates don't need cold stored. They last a long as time sitting on a shelf. The slower they dry out the longer they last.
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NecroMyce
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Quote:
verum subsequentis said: Fresh plates don't need cold stored. They last a long as time sitting on a shelf. The slower they dry out the longer they last.
I’m talking about fresh plates that have not been transferred to, I made a shit load so I don’t have to do it again for a while, just curious what best storage options are.
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: NecroMyce]
#27004832 - 10/26/20 03:01 PM (3 years, 3 months ago) |
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i know
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NecroMyce
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Okay cool. Thank you so much bro! I made a bunch extra, wrapped them Up and put them in plastic bags until further needed!
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: NecroMyce]
#27004932 - 10/26/20 04:03 PM (3 years, 3 months ago) |
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Just no that temperature fluctuations will lead to condensation in the plates and that is not good. If you don't have relatively stable temps I'd suggest putting them in an insulated box to slow down temp swings.
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NecroMyce
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Mine on the shelf will fluctuate 2-6 degrees maybe. But In the fridge it’s stable, that’s why I was asking about cold storage, it’s stable, but not sure if it’ll kill the shelf life.
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: NecroMyce]
#27005071 - 10/26/20 05:21 PM (3 years, 3 months ago) |
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fridges are less stable than you might think. It depends on the quality of the fridge and a few other factors. You're room will be fine.
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verum subsequentis
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I pour 50 to 60 plates at a time, wrap them or put them in ziploc bags and keep them at room temperature. Use as needed.
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Stipe-n Cap


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Quote:
verum subsequentis said: fridges are less stable than you might think. It depends on the quality of the fridge and a few other factors. You're room will be fine.
Definitely, I've lost a lot of decent cultures to iced up plates.
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: Stipe-n Cap]
#27005095 - 10/26/20 05:38 PM (3 years, 3 months ago) |
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Freezerless mini fridges are tits.
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NecroMyce
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Quote:
verum subsequentis said: Freezerless mini fridges are tits.
That’s what I have but once you have your clean master culture, you store in the fridge to stall growth for long term storage?
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: NecroMyce]
#27005113 - 10/26/20 05:50 PM (3 years, 3 months ago) |
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Yes. The cold slows down the growth. The colder the better as long as you don't freeze. Cube myc can't hardly do a thing near freezing. But if you freeze it it'll die.
Slants are best for long time storage but plates will last a good long while if kept properly.
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NecroMyce
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Sweet. I was planning on doing slants eventually, but not until I get my master plates done, Knock up a couple 5qt size grain Growbag, get a few tubs going and see how the fruits do, I plan on cloning a fruit from That first run And then using that plate to make a slant as time goes on, probably sometime next year.
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Phony Phone

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Re: Agar transfer, avoiding isolation [Re: NecroMyce]
#27005873 - 10/27/20 08:19 AM (3 years, 3 months ago) |
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46-48 F (8-9 celsius) is fine fridge temp for storage? OT: Realized the plates i had were unsterilized might explain the satellite contams around the myc. Now must be redone over again
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NecroMyce
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Re: Agar transfer, avoiding isolation [Re: Phony Phone] 1
#27011191 - 10/29/20 09:55 PM (3 years, 2 months ago) |
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I hate to make a thread carry on, but I feel as there is a lot of people learning here. And everyone can learn together, no reason to make a new thread.
So I started PR, CR, AA+, Burma, Malabar all from spore inoculation right on to agar, I made about 3-4 plates for each strain. And of course I had contams, so I transferred those plates. And look at that, most are pretty clean and have rhizoporn growth. Going to let these go a few more days and then make another transfer. Not sure if that’s the best idea, but my plan is to get a few fully grown plates for inoculations. Basically have them in the fridge for whenever I’m ready, a nice clean master culture, once I only have one little piece of the agar left, or one plate left, I’ll use that plate to make more plates and it’s a never ending cycle.
I also clones a few large fruits from my 2nd flush. I plan on letting those go a little longer and doing some transfers and once that’s clean, fruiting those.
Here is some pictures. Not really sure when the best time to transfer is. I’ve read the earlier the better. But I don’t know. I’m also curious if these plates grow out and are perfectly clean, why not just start inoculating with it, and use the last piece to make another master culture. Hmmmm
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: NecroMyce]
#27011620 - 10/30/20 08:01 AM (3 years, 2 months ago) |
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Looks like you're doing wonderfully.
When to transfer just depends on the plate and what's going on. Sometimes you want to hurry and snatch something up before something goes fucky and sometimes you want to wait a minute and watch things grow out so you can see what's up.
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NecroMyce
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But in theory unless we are isolating strains and mainly going for inoculation plates, as long as it’s clean we are good! The idea mainly is to have clean culture to inoculate with? If nothing goes wrong in a plate, let it grow and either store it in a fridge to halt growth, or inoculate some bags? I was gonna drop a decent sized cut from these plates to a 5quart spawn bag to skip the grain to grain process. Am I following?
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Phony Phone

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Re: Agar transfer, avoiding isolation [Re: NecroMyce]
#27012365 - 10/30/20 03:47 PM (3 years, 2 months ago) |
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A kinda trivial question: how do you make your agar? Its pristine
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NecroMyce
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Re: Agar transfer, avoiding isolation [Re: Phony Phone] 1
#27012563 - 10/30/20 06:02 PM (3 years, 2 months ago) |
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PastyWhyte write ups
Here’s a good link,
But here’s the recipe I used D3’s perfect transfers
I know this link above is a link for how to make transfers, but if you read it, it’ll show his agar recipe, a lot of people use Potato flakes and yeast and all that, but I went simple and used just agar and light malt extract. I suggest using the food coloring too because it makes the mycelium nice a white and easier to see contams, and it looks badass! I feel like a science expert, and your family and friends that watch you do agar work or see your progression will think the same lol!
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Phony Phone

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Re: Agar transfer, avoiding isolation [Re: NecroMyce]
#27016713 - 11/02/20 03:56 AM (3 years, 2 months ago) |
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Its me again.
Im uploading 2 pics showing mycelium that seems to be "eaten away" at one point. It's a patch with fuzzier mycelium growth over it. You can see the rhziomorphs stalling right behind the fuzzier growth.
What could it be?
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: Phony Phone]
#27017286 - 11/02/20 11:12 AM (3 years, 2 months ago) |
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Could be ten million different things. Most likely though are the following
A: Contams lurking in the culture and fucking shit up.
B: Just different sets of genetics figuring their shit out and showing different morphology.
C: Some combination of the above.
I'd guess it's just different genetics but it's not easy to tell sometimes. I'd suggest taking transfers and seeing how they do. Looking pretty fuckin good though.
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NecroMyce
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Mr. Mushie

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Re: Agar transfer, avoiding isolation [Re: NecroMyce]
#27027515 - 11/07/20 08:07 PM (3 years, 2 months ago) |
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Man I'm bummed none of my 290 plates all transfer #3 even look close to this good.
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verum subsequentis
seeker of truth



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Re: Agar transfer, avoiding isolation [Re: Mr. Mushie]
#27027674 - 11/07/20 09:56 PM (3 years, 2 months ago) |
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290? What the fuck? Why?
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Phony Phone

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Re: Agar transfer, avoiding isolation [Re: Mr. Mushie]
#27027926 - 11/08/20 04:31 AM (3 years, 2 months ago) |
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pics?
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GrinchGrower
N00B



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Re: Agar transfer, avoiding isolation [Re: Phony Phone]
#27028614 - 11/08/20 01:24 PM (3 years, 2 months ago) |
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Maybe he means 290 total in his lifetime of agar work?
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Mr. Mushie

Registered: 01/16/20
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I understand, it's probably overkill.
The most challenging part of cultivation for me currently is grain prep and agar work. I figured I'm going to lose some jars to contamination along the way so why not make an insane amount of plates so that I can inoculate 100+ jars.

Here's 170 plates all T3 started from two MS syringes. Some have gone to grains, most have not.
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Stipe-n Cap


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Re: Agar transfer, avoiding isolation [Re: Mr. Mushie]
#27028736 - 11/08/20 02:33 PM (3 years, 2 months ago) |
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Quote:
Mr. Mushie said: Here's 170 plates all T3 started from two MS syringes. Some have gone to grains, most have not.
Agar is meant to clean and isolate. Just a tip to consider for the future, if you don't intend to use those plates for creating spawn then having that many is a waste of resources. You aren't helping anything by running that many.
When cleaning just transfer to one or two plates at a time. It doesn't help at all to have that many, even if you're just using in for practice it's still a waste.
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: Mr. Mushie]
#27028746 - 11/08/20 02:38 PM (3 years, 2 months ago) |
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The smartest and most cost and energy effective way to go about that is to make one or two plates of each variety and transfer until you know it's clean. Then you take the clean plate and noc up a few jars. You can make as many or as few jars per dish as you want but i wouldn't recommend more than ten jars per dish. You then let these jars grow out and G2G into more jars. You can turn ten jars into 200 jars in no time. Some cultivators utilized the same principle but choose to expand myc via LC instead. Either way works fine if done properly.
You are wasting time and plates. Look at the wall of plates in my sig. I've been there. That's how i know.
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Mr. Mushie

Registered: 01/16/20
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Thank you both for the response.
My worry is that I'm going to mess up grain prep (too dry, or too wet) and if I only had two plates, that i'd lose all my jars due to noob errors and id be back at the starting line.
I figured making a crazy amount of plates while I practice grain prep with my first 50# bag wouldn't be a terrible idea.
Thanks again for the knowledge verum and p9hu7.
I'm simply worried about a small mistake that ends up ruining 6 weeks of work.
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: Mr. Mushie]
#27028778 - 11/08/20 02:58 PM (3 years, 2 months ago) |
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It's understandable. Still way way overkill. Ten plates would be more than enough. Good news is, if all those plates are good. You can throw extras in the fridge and use as needed. Be sure to take proper precautions if you go the fridge route though.
Grain prep is easy. What type of grain are you using?
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Mr. Mushie

Registered: 01/16/20
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I'm using whole oats.
Tried eats prep and bods prep both a couple times now. Oats were coming out of the PC in one big clump that i'm unable to break up.
Tried to get them completely dry on the outside the last two runs and they're definitely a lot less clumpy/ move more freely, not perfect though.
Going to build a strainer like yours in the coming weeks and use box fans beneath like how lotkid does.
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: Mr. Mushie]
#27029044 - 11/08/20 06:20 PM (3 years, 2 months ago) |
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Yeah. Just hydrate till they are nice and plump and then dry thoroughly. Lack of proper drying is what gets a lot of noobs with oats. I prefer wheat, rye and wbs but oats can work fine.
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mimir
Professional Noob

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Figured this isn't too old to resurrect, but wouldn't a solution possibly be to keep cloning at every grow? If as soon as the mycelium is subcultured on agar it starts to sector and lose genotypes left and right, wouldn't cloning its offspring manage to keep everything together every time? Maybe this could be done once sectoring on agar starts to appear, not necessarily with every single grow.
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Phony Phone

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Re: Agar transfer, avoiding isolation [Re: mimir]
#27177897 - 01/30/21 12:03 PM (2 years, 11 months ago) |
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Yup, but many TC's say that even clones are made up of multiple strains, however i think with a clone you are guaranteed to have a fruiting strain everytime since it came from a mushroom. But from multispore there are so many strains that you'd have to transfer many times in order to get a true isolate. I don't think the average grower is in risk of having that happen to them.
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mimir
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Re: Agar transfer, avoiding isolation [Re: Phony Phone]
#27177907 - 01/30/21 12:17 PM (2 years, 11 months ago) |
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After reading Masters and slaves hunting clones I guess this is more of a theoretical discussion at this point, as keeping a slant + master plate will avoid any excessive subculturing.
Quote:
Yup, but many TC's say that even clones are made up of multiple strains
I never implied this wasn't the case, I think you're confusing with my other post you replied to
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: mimir]
#27179497 - 01/31/21 02:19 PM (2 years, 11 months ago) |
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Cloning again and again will lead to an old and weak culture. Cell division = age. It's why we die eventually (Not that that's all there is to it or that i fully understand life and death). When you start again from spores, you are starting a new baby culture. As it grows across the agar plate/ grain jar/ substrate, it is dividing cells over and over and over.
This is precisely why we slant killer cultures and put them in the fridge. At near freezing temps the culture basically quits growing. By doing this you keep your culture at the age it was when slanted (basically). It's like having a sixteen year old you on tap at any moment.
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mimir
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I am assuming the problem of the culture getting old doesn't happen that easily when isolating on agar. You are still expanding the mycelium but the amount of cell divisions that it takes to get to the cell you are biopsying from the mushroom is much larger than that following even hundreds of transfers. Am I correct in this assumption?
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verum subsequentis
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Re: Agar transfer, avoiding isolation [Re: mimir]
#27181646 - 02/01/21 07:50 PM (2 years, 11 months ago) |
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Yes you are.
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